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目的：建立稳定整合的细胞株，为研究外源基因定位整合机制及其对靶细胞生物特征影响，建立一个生物模型。方法：构建重组病毒载体ＰＬＸＣＤ；用Ｌｉｐｏｆｅｃｔｉｎ 将ＰＬＸＣＤ 转染ＰＡ３１７ 包装细胞，用Ｇ４１８ 筛选获得抗性细胞克隆，扩增抗性细胞收集病毒上清；测定病毒上清效价，感染ＲＰＭＩ８８６６ 细胞，用Ｇ４１８ 筛选获得抗性细胞（ＲＶｃ２３８８６６ 细胞） ；用ＦＡＣＳ 分析ＲＶｃ２３８８６６ 细胞膜蛋白分子ＣＤ２３ 的表达；用细胞原位杂交（ＦＩＳＨ） 分析细胞染色体基因组的改变。结果：建立了稳定整合的ＲＶｃ２３８８６６ 细胞株。结论：ＦＩＳＨ 结果表明ＰＬＸＣＤ 重组体已整合到细胞基因组中；ＦＡＣＳ分析表明ＲＶｃ２３８８６６ 细胞膜表面ＣＤ２３ 分子的表达量明显降低，并说明了整合入基因组的ＣＤ２３ 片段影响了固有ＣＤ２３ 基因的表达。
OBJECTIVE: To establish a stable and stable cell line, and establish a biological model for studying the integration mechanism of exogenous gene localization and its influence on the biological characteristics of target cells. Methods: The recombinant virus vector PLXCD was constructed. The PLXCD cells were transfected with PLXCD by Lipofectin. The resistant cells were screened by G418, and the resistant cells were expanded to collect the virus supernatant. The titer of virus supernatant was assayed and infected with RPMI8866 cells. Screening of resistant cells (RVc23 8 866 cells); FACS analysis RVc23 8866 cell membrane protein CD23 expression; using cell in situ hybridization (FISH) analysis of changes in the cell genome. Results: Stable and integrated RVc238866 cell line was established. Conclusion: The results of FISH showed that the recombinant PLXCD was integrated into the cell genome. FACS analysis showed that the expression of CD23 on the surface of RVc238866 cell membrane was significantly decreased, indicating that the CD23 fragment integrated into the genome affected the expression of intrinsic CD23.